PgsD-677 cells do not produce heparan sulfate. They do produce chondroitin sulfate.

They lack both N-acetylglucosaminyltransferase and glucuronyltransferase activities that are required for synthesis of heparan sulfate.

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53 mMxa0EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
   Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C

Lidholt K, et al. A single mutation affects both N-acetylglucosaminyltransferase and glucuronosyltransferase activities in a Chinese hamster ovary cell mutant defective in heparan sulfate biosynthesis. Proc. Natl. Acad. Sci. USA 89: 2267-2271, 1992. PubMed: 1532254

Esko JD, et al. Tumor formation dependent on proteoglycan biosynthesis. Science 241: 1092-1096, 1988. PubMed: 3137658